ENZYME CONTAINING GRANULATE, METHOD FOR PRODUCTION THEREOF, AND USE THEREOF

Abstract: The enzyme containing granulate is a co-granulate of at least one cellulase and at least one protease. It has been found that the cellulase exhibits a superior stability in this co-granulate, when mixed with a detergent.

Description: ENZYME CONTAINING GRANULATE, METHOD FOR PRODUCTION THEREOF, AND USE THEREOF The invention comprises an enzyme containing granulate, a method for production thereof, and a use thereof. It has been found that a cellulase containing granulate exhibits an unsatisfactory low enzymatic stability under humid conditions if present in a detergent. It has been suggested to coat enzyme containing granulates in order to improve the enzyme stability, vide e.g. WO 89/08695 and US 4,707,287. None of these prior art methods for improvements of the enzyme stability have been satisfactory. Thus, it is the purpose of the invention to provide an enzyme containing granulate, in which the enzyme comprises a cellulase, and in relation to which the cellulase stability under humid conditions in admixture with a detergent is improved. The enzyme containing granulate according to the invention is characterized by the fact that it is a co-granulate comprising an intimate mixture of at least one cellulase and at least one protease, and that it does not contain a photobleaching agent. The cellulases and the proteases used as components in the enzyme containing granulate according to the invention can be produced conventionally, i.e. by means of naturally occurring microorganisms, or by means of protein engineering. It has been found that the granulate according to the invention exhibits a better cellulase stability under humid conditions in admixture with a detergent than the prior art granulates. As it would be assumed a priori that protease might digest the cellulase in the co-granulate and thus impair the cellulase stability, this is a surprising result. When more than one enzyme has to be added to a detergent it is convenient for the detergent manufacturer to add a single granulate containing all the enzymes to be added, i.a. a co-granulate. The most widely used enzyme in detergents is a protease and thus, the skilled worker would anticipate the risk that the protease would decompose the other enzymes during the production of the co- granulate or during storage under humid conditions. The best the skilled worker would anticipate would be that the stability of the other enzymes (e.g. amylases, lipases and/or cellulases) would be unchanged. Most surprisingly, however, as 5 indicated above, it has been found that a cellulase can obtain an improved stability by being co-granulated with a protease. Also, it has been found that the stabilization improving effect related to the granulate according to the invention is not found in a mixture of a cellulase granulate and a protease granulate. Thus, it is imperative for the invention that the 10 granulate is a co-granulate with the composition indicated. A preferred embodiment of the granulate according to the invention is characterized by the fact that the granulate is coated. The granulate can be coated by means of any of the usually used coating agents, e.g. the coating agents described in Australian Patent No. 459399, page 3, lines 4-11. In this manner an 15 improved stability of the cellulase is obtained. A preferred embodiment of the granulate according to the invention is characterized by the fact that the cellulase is one or more of the cellulases producible by means of Humicola insolens DSM 1800, DSM 2068 or DSM 2069 or natural or protein engineered variants thereof. It has been found that these cellulases 20 can be stabilized in an excellent manner. The definition of the cellulase activity unit S-CEVU appears from AF 302. A preferred embodiment of the granulate according to the invention is characterized by the fact that the cellulase is neutral or alkaline. In this context a neutral or alkaline cellulase is a cellulase, the pH optimum of which is between 6 and 25 11 , determined according to AF 302, with the necessary pH adjustsments in order to generate a pH activity curve. These cellulases are preferred because they exhibit the highest activities during their intended use in a wash liquid. A preferred embodiment of the granulate according to the invention is characterized by the fact that the protease is a natural or protein engineered variant 30 of a subtilisin type protease. These proteases are especially well suited as components of enzymatic detergent additives, due to their excellent effect in the washing liquid. A preferred embodiment of the granulate according to the invention is characterized by the fact that the protease is Savinase ® (producible by means of B. lentus), Alcalase ® (producible by means of B. licheniformis) or subtilisin Novo (producible by means of B. subtilis). These proteases exhibit an excellent stabilizing effect. A preferred embodiment of the granulate according to the invention is characterized by the fact that the protease is a natural or protein engineered variant of a trypsin type protease. These proteases exhibit a good stabilizing effect. A preferred embodiment of the granulate according to the invention is characterized by the fact that the protease is producible by means of Fusarium oxysporυm. These proteases exhibit a good stabilizing effect and can be obtained as SP 387 from NOVO NORDISK A/S. A preferred embodiment of the granulate according to the invention is characterized by the fact that the weight proportion between the protease protein and the cellulase protein is at least 0.01, preferably at least 0.1. No satisfactory stabilizing effect has been seen with a weight proportion between the protease protein and the cellulase protein less than 0.01. A preferred embodiment of the granulate according to the invention is characterized by the fact that the granulate also contains PVP, preferably in an amount between 0.001 and 0.1 g/g of granulate. It has been found that the effect of PVP is a further improvement of the cellulase stability under humid conditions in admixture with a detergent. Also, the invention comprises a method for production of the granulate according to the invention, which is characterized by the fact that an aqueous solution containing at least one of the enzymes cellulase and protease is sprayed onto a solid core in a granulating equipment. It is to be understood that the core shall contain a protease, if the aqueous solution contains a cellulase only, and vice versa. Also, it is to be understood that the core may be enzyme free, if the aqueous solution contains both a cellulase and a protease. Also, the invention comprises a use of the granulate according to the • invention, as a detergent additive. Thus, the granulate according to the invention can be used as a detergent additive in any detergent, e.g. both detergents with and without oxidizing agents. A preferred embodiment of the use according to the invention is characterized by the fact that the granulate is added in an amount of between 0.1 and 5% w/w, related to the total detergent weight. This corresponds to the concentrations of enzymatic additive in a commercial detergent. WO 90/15856 describes a co-granulate, which among many possibilities may contain both a cellulase and a protease. This prior art co-granulate, however, always contains a photobleaching agent and thus differs significantly from the invention. Danish Patent No. 157937 describes a granulate with a core and a shell, in which the core may contain a cellulase and the shell may contain a protease. However, this granulate is not a co-granulate, and it does not comprise the intimate mixture of the cellulase and the protease, which is found in the granulate according to the invention. Also, it has been found that the cellulase stability of this prior art granulate is inferior in comparison to the cellulase stability of the granulate according to the invention. Documentation for this will be presented later in this specification. WO 90/09440 describes a granulate with a core and a shell, in which the core contains one enzyme and the shell contains another enzyme. However, this granulate is not a co-granulate, it contains a sustained release coating between the core and the shell, and it does not comprise the intimate mixture of the two enzymes which is found in the granulate according to the invention. Accordingly, it has been found that the cellulase stability of this prior art granulate is inferior in comparison to the cellulase stability of the granulate according to the invention. The invention will be illustrated by the following examples. In order to demonstrate the stability improving effect provided by means of the invention directly, some of the examples are examples illustrating the invention, i.e. Examples 1, 2, 6, 7, 9, 12, 15, 16, 17, 19, and 20, and some of the examples are comparison examples illustrating the prior art, i.e. Examples 3, 4, 5, 8, 10, 11 , 13, 14, and 18. The cellulase activity S-CEVU is measured according to AF 302; the protease activity AU is measured according to AF 218, the protease activity KNPU is measured according to AF 220, the amylase activity KNU is measured according to AF 214, and the lipase activity KLU is measured according to AF 95. EXAMPLE 1 The following powder components are used for the production of 15 kg of granulate: 2.50 kg of ground Alcalase ® concentrate (14.6 AU/g) 1.50 kg of fibrous cellulose ARBOCEL BC 200 8.70 kg of finely ground sodium sulphate 0.60 kg of calcium carbonate 0.60 kg of dextrin The above indicated components are mixed in a 501 Lόdige mixer, and simultaneously the temperature is raised to 34°C. The mixing time is 2 minutes, the velocity of the mixture paddles is 145 rpm, and the velocity of the chopper knives is 3000 rpm. With the same velocities as above indicated 3.3 kg of an enzyme mixture consisting of 1.00 kg of liquid cellulase concentrate (45,000 S-CEVU/g, 31% dry matter) 1.50 kg of water 0.60 kg of lactose 0.23 kg of PVP is sprayed onto the above indicated mixed powder components, and subsequently 1.2 kg of water is sprayed thereon. A pneumatic spraying nozzle is used, and the spraying time is around 6 minutes. After the spraying the wet mixture is granulated further for 3 minutes, and a sphere or lense shaped granulate is formed. The humid granulate is dried at 60°C to a water content of less than 3%. The dry granulate is sieved to a particle size range between 300 and 1000 μm: Fines < 300 μm: 0.81 kg Agglomerates > 1000 μm: 0.40 kg Wanted product 300-1000 μm: 8.30 kg Activity: Cellulase 3200 S-CEVU/g Protease 2.46 AU/g A part of the wanted product is coated in the following manner. 2 kg of the wanted product is heated to 75-80°C in a 5 litres heat jacketed Lόdige mixer, and this temperature is maintained during the entire coating process. The heated granulate is mixed with 100 g of melted PEG 4000 followed by a powdering with 250 g of Ti0 2 and a subsequent addition of 20 g of PEG 4000. The granulate is finally cooled to room temperature in a fluid bed and siftet to a particle size range of 300-1000 μm. The obtained granulate exhibited a weight proportion between Alcalase ® protein and cellulase protein of approximately 4.4, a cellulase activity of 2690 S-CEVU/g, and a protease activity of 2.0 AU/g. EXAMPLE 2 The following powder components are used for the production of 15 kg of granulate 11.5% of fibrous cellulose, ARBOCEL BC 200 4.0% of kaolin, SPESWHITE ECC 60.0% of finely ground sodium sulphate 10.0% of carbohydrate binder 5 11.0% of Savinase ® concentrate (34.8 KNPU/g) 1.0% of polyvinyl pyrrolidone (PVP) KOLIDON K-30 The above indicated components are granulated in a FM 50 Lόdige mixer as in Example 1. Also, as in Example 1 lactose (only 50% of the amount used in Example 1, though) and PVP is dissolved in a mixture of liquid cellulase 10 concentrate and water. The thus formed granulate is dried in a fluid bed to a water content of below 3%, and subsequently sieved in order to obtain a product with a particle size range from 300 to 1000 μm. A part of the wanted product is coated in the following manner. 2 kg 15.of the wanted product is heated to 75-80°C in a 5 litres heat jacketed Lόdige mixer, and this temperature is maintained during the entire coating process. The heated granulate is mixed with 100 g of melted PEG 4000 followed by a powdering with 250 g of Ti0 2 and a subsequent addition of 20 g of PEG 4000. The granulate is finally cooled to room temperature in a fluid bed and 0 siftet to a particle size range of 300-1000 μm. Activity: Cellulase 1650 S-CEVU/g Protease 3.30 KNPU/g The obtained granulate exhibited a weight proportion between Savinase ® protein and cellulase protein of approximately 2.1. 5 In the following documentation is presented for the improvement of the cellulase stability in the co-granulate according to the invention in the presence of a detergent. As a reference a blend of a cellulase granulate and a protease granulate in the presence of a detergent has been used. The experimental conditions were as follows. The granulate according to the invention to be tested (Examples 1 and 2) is weighed into a series of glasses with approx. 3200 S-CEVU/glass. 8.0 g of a detergent, which is a compact zeolite built detergent without bleach is then added to each glass, and the glasses are closed with a lid. Subsequently, the content of the closed glasses is mixed by shaking. The lids are removed, and then the glasses are introduced into a climate room at 80% relative humidity and 35°C. After storage for a certain number of days, the whole content of each glass is analyzed for cellulase activity. The prior art granulates to be tested are blends of granulates each of which contains only one enzyme component. Then granulates are produced as indicated in Example 2 with the exception that only one enzyme is added. Thus, in case a Savinase granulate is to be produced, the cellulase is omitted and replaced by approximately the same amount of water, and in case a cellulase granulate is to be produced, the Savinase is omitted and replaced by the same amount of sodium sulphate. The results were as follows. It appears from the above that the stability of the cellulase in the co- granulate is considerably improved in comparison to the stability of the cellulase in the prior art granulates. EXAMPLE 3 This example and Example 4 are not examples according to the invention because no protease is present in the list of components. Instead skim milk powder is used in order to investigate, if skim milk powder showed any stability improving effect on the cellulase. A powder mixture with the following composition: 1.50 kg fibrous cellulose, ARBOCEL BC 200 0.60 kg of CaC0 3 CALOFORT type L 11.80 kg finely ground sodium sulphate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.9 kg of a granulating liquid made up by mixing 1.20 kg skim milk powder 0.15 kg PVP KOLIDON K30 3.55 kg aqueous Carezyme ® cellulase concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between casein protein and cellulase protein of approximately 7.9 and a cellulase activity of 971 S- CEVU/g. EXAMPLE 4 A powder mixture with the following composition: 1.70 kg fibrous cellulose, ARBOCEL BC 200 0.64 kg of CaC0 3 CALOFORT type L 8.60 kg finely ground sodium sulphate 3.80 kg skim milk powder 5 is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 3.6 kg of a granulating liquid made up by mixing 0.16 kg PVP KOLIDON K30 3.44 kg aqueous Carezyme ® concentrate Carezyme ® is a cellulase preparation produced according to WO 10 91/17243. The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between casein protein and cellulase protein of approximately 15 and a cellulase activity of 1600 S- CEVU/g. 15 EXAMPLE 5 This example is not an example according to the invention because no protease is present in the list of components. Instead gelatine is used in order to investigate, if gelatine showed any stability improving effect on the cellulase. A powder mixture with the following composition: 0 1.50 kg fibrous cellulose, ARBOCEL BC 200 0.60 kg of CaC0 3 CALOFORT type L 12.00 kg finely ground sodium sulphate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.1 kg of a granulating liquid heated to approximately 40°C made up by mixing 0.60 kg carbohydrate binder 0.15 kg Gelatine Bloom 200 3.55 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between gelatine protein and cellulase protein of approximately 2.3 and a cellulase activity of 1096 S- CEVU/g. O The reference is identical to Example 8, but with Na 2 S0 4 instead of BAN EXAMPLE 6 A powder mixture with the following composition: 1.70 kg fibrous cellulose, ARBOCEL BC 200 0.64 kg of CaC0 3 CALOFORT type L 9.60 kg finely ground sodium sulphate 0.64 kg carbohydrate binder 1.70 kg Alcalase ® concentrate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 3.9 kg of a granulating liquid made up by mixing 0.16 kg PVP KOLIDON K30 0.40 kg carbohydrate binder 5 3.44 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between Alcalase ® protein and cellulase protein of approximately 2.3, a cellulase activity of 10 1790 S-CEVU/g and a protease activity of 0.867 AU/g. ~ ) The reference is identical to Example 8, but with Na;,S0 4 instead of BAN 15 EXAMPLE 7 A powder mixture with the following composition: 1.78 kg fibrous cellulose, ARBOCEL BC 200 0.64 kg kaolin 10.50 kg finely ground sodium sulphate 0 0.48 kg carbohydrate binder 0.80 kg Savinase ® concentrate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.2 kg of a granulating liquid made up by mixing 0.24 kg PVP KOLIDON K30 0.96 kg carbohydrate binder 3.30 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between protease protein and cellulase protein of approximately 0.32, a cellulase activity of 3900 S- CEVU/g and a protease activity of 1.14 KNPU/g. EXAMPLE 8 This example is not an example according to the invention because no protease is present in the list of components. Instead amylase is used in order to investigate, if amylase showed any stability improving effect on the cellulase. A powder mixture with the following composition: 1.78 kg fibrous cellulose, ARBOCEL BC 200 1.28 kg kaolin 9.90 kg finely ground sodium sulphate 0.64 kg carbohydrate binder is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.2 kg of a granulating liquid made up by mixing 0.24 kg PVP ' KOLIDON K30 0.64 kg carbohydrate binder 1.30 kg Amylase BAN concentrate 2.00 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between amylase protein and cellulase protein of approximately 0.16, a cellulase activity of 4790 S- CEVU/g and an amylase activity of 48.8 KNU/g. EXAMPLE 9 A powder mixture with the following composition: 1.78 kg fibrous cellulose, ARBOCEL BC 200 0.64 kg kaolin 10.00 kg finely ground sodium sulphate 0.48 kg carbohydrate binder 0.80 kg Alcalase ® concentrate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.5 kg of a granulating liquid made up by mixing 0.24 kg PVP KOLIDON K30 1.44 kg carbohydrate binder 2.80 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between protease protein and cellulase protein of approximately 0.43, a cellulase activity of 4900 S- CEVU/g and a protease activity of 0.305 AU/g. k ) The reference is identical to Example 8, but with Na 2 S0 4 instead of BAN EXAMPLE 10 A reference T-granulate is produced as described in Example 1 but without protease. This PEG 4000 coated granulate is further spray coated on the outside of the granules with Savinase ® in a fluid bed. The obtained granulate exhibits a weight proportion between protease protein and cellulase protein of approximately 0.7, a cellulase acitivty of 5100 S- CEVU/g, and a protease activity of 3.5 KNPU/g. The obtained granulate is a co-granulate comprising a cellulase and a protease, but the cellulase and protease are not intimately mixed, becauase the cellulase is in the core and the protease (Savinase ® ) is in the coating. For that reason the granulate is a reference granulate, as above indicated. EXAMPLE 11 The granulate is produced as in Example 10, except for the fact that twice the amount of Savinase ® is coated on the outside of the granules. The obtained granulate exhibited a weight proportion between protease protein and cellulase protein of approximately 1.4, a cellulase activity of 4800 S- CEVU/g, and a protease activity of 6.9 KNPU/g. EXAMPLE 12 A granulate was produced as described in Example 1 , except for the fact that the amounts of cellulase, water and protease are adjusted to generate the following activities: 618 S-CEVU/g (cellulase) and 3.19 AU/g (protease). The weight proportion between protease protein and cellulase protein was approximately 30. *) The reference is identical to Example 8, but with Na,S0 4 instead of BAN EXAMPLE 13 This example is not an example according to the invention because no protease is present in the list of components. Instead Termamyl ® (an amylase) is used in order to investigate, if this amylase showed any stability improving effect on the cellulase. A powder mixture with the following composition: 1.78 kg fibrous cellulose, ARBOCEL BC 200 1.28 kg kaolin 10.90 kg finely ground sodium sulphate 5 0.80 kg Termamyl ® concentrate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.3 kg of a granulating liquid made up by mixing 0.64 kg carbohydrate binder 3.66 kg aqueous Carezyme ® concentrate o The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between amylase protein and cellulase protein of approximately 0.60, a cellulase activity of 4700 S- CEVU/g and an amylase activity of 60 KNU/g. 5 EXAMPLE 14 This example is not an example according to the invention because no protease is present in the list of components. Instead Lipolase ® (a lipase) is used in order to investigate, if this lipase showed any stability improving effect on the cellulase. A powder mixture with the following composition: 1.78 kg fibrous cellulose, ARBOCEL BC 200 1.28 kg kaolin 11.30 kg finely ground sodium sulphate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.7 kg of a granulating liquid made up by mixing 0.64 kg carbohydrate binder 1.50 kg Lipolase ® concentrate 2.60 kg aqueous Carezyme ® concentrate The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. The obtained granulate exhibited a weight proportion between lipase protein and cellulase protein of approximately 0.51 , a cellulase activity of 5060 S- CEVU/g and a lipase activity of 35 KLU/g. *) The reference is identical to Example 8, but with NagS0 4 instead of BAN **) This example was not performed concurrently with the three above examples, but is listed for comparison purposes It appears from the foregoing that the cellulase stability improving effect is a selective protease effect, because other enzymes than protease and other proteins do not generate the cellulase stability improving effect. Also, it appears that the co-granulate according to the invention necessarily has to comprise an intimate mixture of cellulase and ' protease in order to generate the cellulase stability improving effect, vide Example 10. EXAMPLE 15 A powder mixture with the following composition: 1.92 kg fibrous cellulose, ARBOCEL BC 200 0.96 kg kaolin 9.80 kg finely ground sodium sulphate 0.32 kg yellow dextrin 1.60 kg Esperase ® concentrate is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.5 kg of a granulating liquid made up by mixing 0.70 kg carbohydrate binder 3.80 kg aqueous Carezyme ® cellulase concentrate (22% DS) The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. A granulate with an activity of 5900 S-CEVU/g and 1.80 KNPU/g was produced. The weight proportion between protease protein and cellulase protein was approximately 0.44. EXAMPLE 16 A powder mixture with the following composition: 1.92 kg fibrous cellulose, ARBOCEL BC 200 0.96 kg kaolin 11.30 kg finely ground sodium sulphate 0.32 kg yellow dextrin is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.2 kg of a granulating liquid made up by mixing 0.58 kg carbohydrate binder 2.72 kg Carezyme ® cellulase concentrate (DS = 32%) 0.90 kg liquid Alcalase ® protease concentrate (DS = 16%) The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. A granulate with an activity of 4300 S-CEVU/g and 0.14 AU/g was produced. The weight proportion between protease protein and cellulase protein was approximately 0.19. ") The reference is identical to Example 8, but with Na 2 S0 4 instead of BAN EXAMPLE 17 A granulate is produced as described in Example 15, but Durazyme ® protease is used instead of Esperase ® protease. At the same time is used 11.1 kg sodium sulfate and 0.68 kg kaolin. A granulate with an activity of 4200 S-CEVU/g and 2.6 DPU/g was produced. *) The reference is identical to Example 8, but with Na j SO,, instead of BAN EXAMPLE 18 A powder mixture with the following composition: 1.80 kg fibrous cellulose, ARBOCEL BC 200 0.60 kg kaolin 10.40 kg finely ground sodium sulphate 0.45 kg yellow dextrin is transferred to a 50 liter Lόdige mixer FM 50 and granulated with 4.4 kg of a granulating liquid made up by mixing 0.80 kg carbohydrate binder 3.60 kg Endolase ® cellulase concentrate (34% DS) The granulation is carried out as described in Example 1. The granulate is dried and sifted and finally coated as described in Example 2. A granulate with an activity of 2300 S-CEVU/g was produced. EXAMPLE 19 This granulate is produced as described in Example 18 but to the powder mixture is added 0.40 kg Alcalase ® powder concentrate instead of 0.40 kg sodium sulfate. A granulate with an activity of 1910 S-CEVU/g and 0.22 AU/g was produced. EXAMPLE 20 This granulate is produced as described in Example 18, but the liquid feed is made of 3.2 kg Endolase ® cellulase concentrate 0.7 kg Alcalase ® concentrate 0.7 kg carbohydrate binder A granulate with an activity of 2300 S-CEVU/g and 0.12 AU/g was produced.

Claims: CLAIMS 1. Enzyme containing granulate, characterized by the fact that it is a co- granulate comprising an intimate mixture of at least one cellulase and at least one protease, and that it does not contain a photobleaching agent. 2. Granulate according to Claim 1, characterized by the fact that the granulate is coated. 3. Granulate according to Claims 1 - 2, characterized by the fact that the cellulase is one or more of the cellulases producible by means of Humicola insolens DSM 1800, DSM 2068 or DSM 2069 or natural or protein engineered variants thereof. 4. Granulate according to Claims 1 - 3, characterized by the fact that the cellulase is neutral or alkaline. 5. Granulate according to Claims 1 - 4, characterized by the fact that the protease is a natural or protein engineered variant of a subtilisin type protease. 6. Granulate according to Claims 1 - 4, characterized by the fact that the protease is Savinase ® (producible by means of B. lentus), Alcalase ® (producible by means of B. licheniformis) or subtilisin Novo (producible by means of B. subtilis). 7. Granulate according to Claims 1 - 4, characterized by the fact that the protease is a natural or protein engineered variant of a trypsin type protease. 8. Granulate according to Claims 1 - 4, characterized by the fact that the protease is producible by means of Fusarium oxysporum. 9. Granulate according to Claims 1 - 8, characterized by the fact that the weight proportion between the protease protein and the cellulase protein is at least 0.01 , preferably at least 0.1. 10. Granulate according to Claims 1 - 9, characterized by the fact that the granulate also contains PVP, preferably in an amount between 0.001 and 0.1 g/g of granulate. 11. Method for production of the granulate according to Claims 1 - 10, characterized by the fact that an aqueous solution containing at least one of the enzymes cellulase and protease is sprayed onto a solid core in a granulating equipment. 12. Use of the granulate according to Claims 1 - 10, as a detergent additive. 13. Use according to Claim 12, characterized by the fact that the granulate is added in an amount of between 0.1 and 5% w/w, related to the total detergent weight.

Inventors: JENSEN POUL ERIK, LAURIDSEN CARSTEN, JENSEN, POUL, ERIK, LAURIDSEN, CARSTEN
Applicants: JENSEN POUL ERIK, LAURIDSEN CARSTEN, NOVONORDISK AS, JENSEN, POUL, ERIK, LAURIDSEN, CARSTEN, NOVO NORDISK A/S
Application year: 1994
Main classification: C12N9
IPC codes: C11D3, C12N9